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Am J Physiol Lung Cell Mol Physiol (October 14, 2005). doi:10.1152/ajplung.00351.2005
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Submitted on August 11, 2005
Accepted on October 5, 2005

Membrane capacitance and conductance changes parallel mucin secretion in the human airway epithelium

Henry Danahay1, Hazel C Atherton1, Alan D Jackson1, James L Kreindler2*, Christopher T Poll1, and Robert J Bridges3

1 Novartis Respiratory Research Centre, Horsham, West Sussex, United Kingdom
2 Department of Pediatrics, Children's Hospital of Pittsburgh of UPMC, Pittsburgh, PA, USA
3 Department of Physiology and Biophysics, Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA

* To whom correspondence should be addressed. E-mail: james.kreindler{at}chp.edu.

The measurement of the magnitude and kinetics of exocytosis from intact epithelia has historically been difficult to perform. Using well differentiated cultures of human bronchial epithelial cells (HBEs) we describe the use of transepithelial impedance analysis to enable the real time quantification of mucin-secretagogue-induced changes in membrane capacitances (surface area) and conductances. ATP{gamma}S, UTP, ionomycin and PMA all induced robust increases in total cellular capacitance that were demonstrated to be dominated by a specific increase in apical membrane surface area. The UTP-induced increase in capacitance occurred in parallel with goblet cell emptying and the secretion of mucin and was associated with decreases in apical and basolateral membrane resistances. The magnitude and kinetics of the capacitance increases were dependent upon both the agonist used and the sidedness of stimulation. The peak increase in capacitance induced by UTP equated to ~30 mucin granule fusions per goblet cell. Secretagogue-induced decreases in apical membrane resistance were independent of exocytosis, although each of the secretagogues induced profound reductions in the basolateral membrane resistance. Transepithelial impedance analysis offers the potential to study morphological and conductance changes in cultured HBEs.




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